Mufazalov Ilgiz A, Waisman Ari
Institute for Molecular Medicine, University Medical Center of the Johannes Gutenberg-University of Mainz, Obere Zahlbacher Str. 67, 55131, Mainz, Germany,
Methods Mol Biol. 2016;1304:73-9. doi: 10.1007/7651_2014_114.
Leukocyte infiltration of the central nervous system (CNS) occurs under certain pathogenic conditions and most often results in severe disorders. Therefore, the isolation and analysis of such infiltrating cell populations is necessary for elucidating the underlying pathogenic mechanisms. Here we describe a simple and straightforward protocol for cell isolation from the inflamed CNS, which combines mechanical dissociation and enzymatic degradation of the tissue. Additionally, purification by Percoll gradient centrifugation provides a great yield of the infiltrating material. The isolated cells can be further used for downstream applications such as cell sorting, cellular or molecular analysis.
中枢神经系统（CNS）的白细胞浸润在某些致病条件下会发生，且最常导致严重疾病。因此，分离和分析此类浸润细胞群体对于阐明潜在的致病机制是必要的。在此，我们描述了一种从炎症性中枢神经系统分离细胞的简单直接的方案，该方案结合了组织的机械解离和酶促降解。此外，通过Percoll梯度离心进行纯化可大量获得浸润物质。分离出的细胞可进一步用于下游应用，如细胞分选、细胞或分子分析。
